Cloned (Comment) | Organism |
---|---|
gene dhaT, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain JM109 | Levilactobacillus brevis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | slight inhibition at 5 mM | Levilactobacillus brevis | |
EDTA | 91% inhibition at 5 mM | Levilactobacillus brevis | |
PMSF | 85% inhibition at 5 mM | Levilactobacillus brevis | |
Zn2+ | slight inhibition at 5 mM | Levilactobacillus brevis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0116 | - |
NADH | pH 7.5, 37°C | Levilactobacillus brevis | |
0.0904 | - |
NAD+ | pH 9.5, 25°C | Levilactobacillus brevis | |
1.25 | - |
3-hydroxypropanal | pH 7.5, 37°C | Levilactobacillus brevis | |
2.26 | - |
Propane-1,3-diol | pH 9.5, 25°C | Levilactobacillus brevis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | activates to 150% at 5 mM, the enzyme contains two Fe2+-binding motifs, amino acid residues Asp189, Gln193, His258, and His272, are involved in the Fe2+ binding | Levilactobacillus brevis | |
Mn2+ | activates to 280% at 5 mM | Levilactobacillus brevis | |
additional information | addition of Co2+, Ni2+, or Mg2+ do not significantly affect PDOR activity at 5 mM | Levilactobacillus brevis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
42000 | - |
- |
Levilactobacillus brevis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-hydroxypropanal + NADH + H+ | Levilactobacillus brevis | reduction of the aldehyde is the preferred reaction | propane-1,3-diol + NAD+ | - |
r | |
3-hydroxypropanal + NADH + H+ | Levilactobacillus brevis 6239 | reduction of the aldehyde is the preferred reaction | propane-1,3-diol + NAD+ | - |
r | |
propane-1,3-diol + NAD+ | Levilactobacillus brevis | - |
3-hydroxypropanal + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Levilactobacillus brevis | A0A0C1Q6R1 | - |
- |
Levilactobacillus brevis 6239 | A0A0C1Q6R1 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme 7.1-7.3fold from Escherichia coli strain JM109 by nickel affinity chromatography and gel filtration | Levilactobacillus brevis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
32.6 | - |
purified recombinant His-tagged enzyme, substrate propane-1,3-diol, pH 9.5, 25°C | Levilactobacillus brevis |
52.7 | - |
purified recombinant His-tagged enzyme, substrate 3-hydroxypropanal, pH 7.5, 37°C | Levilactobacillus brevis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,4-butanediol + NAD+ | - |
Levilactobacillus brevis | 4-hydroxybutanal + NADH + H+ | - |
r | |
1-butanol + NAD+ | - |
Levilactobacillus brevis | 1-butanal + NADH + H+ | - |
r | |
1-propanol + NAD+ | - |
Levilactobacillus brevis | 1-propanal + NADH + H+ | - |
r | |
3-hydroxypropanal + NADH + H+ | reduction of the aldehyde is the preferred reaction | Levilactobacillus brevis | propane-1,3-diol + NAD+ | - |
r | |
3-hydroxypropanal + NADH + H+ | reduction of the aldehyde is the preferred reaction, 3-hydroxypropanal is the preferred substrate | Levilactobacillus brevis | propane-1,3-diol + NAD+ | - |
r | |
3-hydroxypropanal + NADH + H+ | reduction of the aldehyde is the preferred reaction | Levilactobacillus brevis 6239 | propane-1,3-diol + NAD+ | - |
r | |
3-hydroxypropanal + NADH + H+ | reduction of the aldehyde is the preferred reaction, 3-hydroxypropanal is the preferred substrate | Levilactobacillus brevis 6239 | propane-1,3-diol + NAD+ | - |
r | |
acetone + NADH + H+ | - |
Levilactobacillus brevis | ? + NAD+ | - |
r | |
dihydroxyacetone + NADH + H+ | - |
Levilactobacillus brevis | ? + NAD+ | - |
r | |
ethanol + NAD+ | - |
Levilactobacillus brevis | acetaldehyde + NADH + H+ | - |
r | |
ethanol + NAD+ | - |
Levilactobacillus brevis 6239 | acetaldehyde + NADH + H+ | - |
r | |
glyceraldehyde + NADH + H+ | - |
Levilactobacillus brevis | ? + NAD+ | - |
r | |
glycerol + NAD+ | - |
Levilactobacillus brevis | dihydroxyacetone + NADH + H+ | - |
r | |
glycerol + NAD+ | - |
Levilactobacillus brevis 6239 | dihydroxyacetone + NADH + H+ | - |
r | |
hydroxyacetone + NADH + H+ | - |
Levilactobacillus brevis | ? + NAD+ | - |
r | |
additional information | the enzyme shows a broad substrate specificity, PDOR can help reduce a broad range of aldehydes and ketones including 3-HPA, propionaldehyde, glyceraldehyde, acetone, hydroxyacetone, and dihydroxyacetone. No activity with acrolein. PDOR can also help oxidize many kinds of alcohols to generate the corresponding aldehydes, and this enzyme is most active with diols containing two primary hydroxy groups separated by one or two carbon atoms. Structure modeling, overview | Levilactobacillus brevis | ? | - |
? | |
additional information | the enzyme shows a broad substrate specificity, PDOR can help reduce a broad range of aldehydes and ketones including 3-HPA, propionaldehyde, glyceraldehyde, acetone, hydroxyacetone, and dihydroxyacetone. No activity with acrolein. PDOR can also help oxidize many kinds of alcohols to generate the corresponding aldehydes, and this enzyme is most active with diols containing two primary hydroxy groups separated by one or two carbon atoms. Structure modeling, overview | Levilactobacillus brevis 6239 | ? | - |
? | |
propane-1,2-diol + NAD+ | - |
Levilactobacillus brevis | 2-hydroxypropanal + NADH + H+ | - |
r | |
propane-1,3-diol + NAD+ | - |
Levilactobacillus brevis | 3-hydroxypropanal + NADH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 42500, recombinant His-tagged enzyme, SDS-PAGE, x * 41993, sequence calculation | Levilactobacillus brevis |
Synonyms | Comment | Organism |
---|---|---|
DhaT | - |
Levilactobacillus brevis |
PDOR | - |
Levilactobacillus brevis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
alcohol oxidation | Levilactobacillus brevis |
37 | - |
aldehyde reduction | Levilactobacillus brevis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | 40 | activity range, aldehyde reduction, profile overview | Levilactobacillus brevis |
25 | 45 | activity range, alcohol oxidation, profile overview | Levilactobacillus brevis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | 45 | purified recombinant His-tagged enzyme, 50% activity remaining after 3 h and 30% after 5 h at 30-37°C, inactivation after 2 h at 45°C | Levilactobacillus brevis |
37 | - |
60% of enzyme relative activity remain after a 2h incubation at 37°C | Levilactobacillus brevis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
aldehyde reduction | Levilactobacillus brevis |
9.5 | - |
alcohol oxidation | Levilactobacillus brevis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 10 | activity range, aldehyde reduction, profile overview | Levilactobacillus brevis |
8 | 10.5 | activity range, alcohol oxidation, profile overview | Levilactobacillus brevis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
PDOR is more stable in acid buffer than in alkaline condition | Levilactobacillus brevis |
6 | 10 | purified recombinant His-tagged enzyme, 50% activityy remaining after 1 h and inactivation aafter 5 h at pH 6.0, 50% activity remaining after 3 h and 30% after 5 h at pH 7.5, inactivation after 1 h at pH 10.0 | Levilactobacillus brevis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | the enzyme contains a cofactor motif, the conserved sequence G-GG-S-X-X-D. The binding site for cofactor NAD(H) is located in the deep hydrophilic pocket between the PDOR two domains | Levilactobacillus brevis | |
NADH | - |
Levilactobacillus brevis |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the type III alcohol dehydrogenases | Levilactobacillus brevis |
additional information | the active site of PDOR is composed of the following amino acid residues, Asp32, Gly92, Gly93, Ser94, Thr134, Thr135, Thr138, Val146, Lys155, Leu177, Asp189, Leu182, Gln193, His258, and His272, which include the binding sites of Fe2+ and the cofactor NAD(H) | Levilactobacillus brevis |